MicroRNA-210 modulates endothelial cell response to hypoxia and inhibits the receptor tyrosine kinase ligand Ephrin-A3
MicroRNA-210 modulates endothelial cell response to hypoxia and inhibits the receptor tyrosine kinase ligand Ephrin-A3
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DOI：
10.1074/jbc.m800731200
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发表时间：
2008-06-06
影响因子：
4.8
通讯作者：
Martelli, Fabio
中科院分区：
文献类型：
--
作者：
Fasanaro, Pasquale;D'Alessandra, Yuri;Martelli, Fabio
microRNAs（miRNAs）是一类非蛋白质编码的小分子RNA，具有负调控基因表达的功能。在本研究中，我们研究了miRNAs在内皮细胞缺氧反应中的作用。我们发现miR-210的表达在暴露于缺氧后逐渐增加。常氧内皮细胞中miR-210过表达刺激Matrigel上毛细血管样结构的形成和血管内皮生长因子驱动的细胞迁移。相反，通过抗-miRNA转染的miR-210阻断抑制了缺氧刺激的毛细血管样结构的形成，并减少了对血管内皮生长因子的反应的细胞迁移。miR-210过表达在常氧和缺氧条件下均不影响内皮细胞生长。然而，antimiR-210转染抑制细胞生长和诱导凋亡，在常氧和缺氧。我们确定了miR-210在缺氧中的一个相关靶点是Ephrin-A3，因为miR-210是下调其表达所必需且足够的。此外，荧光素酶报告基因测定显示Ephrin-A3是miR-210的直接靶点。通过miR-210调节Ephrin-A3具有显著的功能性后果;事实上，不被miR-210靶向的Ephrin-A3等位基因的表达阻止了miR-210介导的对微管发生和趋化性的刺激。我们的结论是，miR-210上调是内皮细胞对缺氧反应的关键因素，影响细胞存活、迁移和分化。
MicroRNAs (miRNAs) are small non-protein-coding RNAs that function as negative gene expression regulators. In the present study, we investigated miRNAs role in endothelial cell response to hypoxia. We found that the expression of miR-210 progressively increased upon exposure to hypoxia. miR-210 overexpression in normoxic endothelial cells stimulated the formation of capillary-like structures on Matrigel and vascular endothelial growth factor-driven cell migration. Conversely, miR-210 blockade via anti-miRNA transfection inhibited the formation of capillary-like structures stimulated by hypoxia and decreased cell migration in response to vascular endothelial growth factor. miR-210 overexpression did not affect endothelial cell growth in both normoxia and hypoxia. However, antimiR-210 transfection inhibited cell growth and induced apoptosis, in both normoxia and hypoxia. We determined that one relevant target of miR-210 in hypoxia was Ephrin-A3 since miR-210 was necessary and sufficient to down-modulate its expression. Moreover, luciferase reporter assays showed that Ephrin-A3 was a direct target of miR-210. Ephrin-A3 modulation by miR-210 had significant functional consequences; indeed, the expression of an Ephrin-A3 allele that is not targeted by miR-210 prevented miR-210-mediated stimulation of both tubulogenesis and chemotaxis. We conclude that miR-210 up-regulation is a crucial element of endothelial cell response to hypoxia, affecting cell survival, migration, and differentiation.